Isolation and mapping of 62 new RFLP markers on human chromosome 11.
Isolation and mapping of 62 new RFLP markers on human chromosome 11.
复制标题
人类 11 号染色体上 62 个新 RFLP 标记的分离和定位。
DOI：
10.1182/blood.v88.4.1177.bloodjournal8841177
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发表时间：
1991
影响因子：
9.8
通讯作者：
Yusuke Nakamura
中科院分区：
文献类型：
--
作者：
T. Tokino;E. Takahashi;M. Mori;A. Tanigami;T. Glaser;T. Glaser;J. W. Park;Carol Jones;T. Hori;Yusuke Nakamura
为了获得人类11号染色体上的新的RFLP标记，以获得高分辨率的图谱，我们从中国仓鼠×人体细胞杂交细胞系中构建了一个粘粒文库，该文库在中国仓鼠的基因组背景中只保留了人11号染色体。通过与标记的人类基因组DNA进行菌落杂交，共分离出3500个宇宙体。从130个粘粒克隆中提取DNA，并进行RFLP检测。在62个RFLP中，检测到一个或多个酶的多态性；4个RFLP为VNTR系统。所有的多态克隆都被分配到22个区间中的一个，通过在包含部分11号染色体的15个体细胞杂交细胞系的缺失小组上进行定位，11个克隆通过原位杂交得到了精细的定位。虽然RFLP标记散布在整个染色体上，但它们主要分布在R-带区域。这些DNA标记将有助于精细定位位于11号染色体上的遗传性疾病基因和肿瘤抑制基因。此外，由于三分之一的粘粒克隆在中国仓鼠DNA中显示了一条或多条带，表明序列保守，这一克隆子集可能有助于分离11号染色体上具有生物重要性的基因。
To obtain new RFLP markers on human chromosome 11 for a high-resolution map, we constructed a cosmid library from a Chinese hamster x human somatic hybrid cell line that retains only human chromosome 11 in a Chinese hamster genomic background. A total of 3,500 cosmids were isolated by colony hybridization with labeled human genomic DNA. DNA was prepared from 130 of these cosmid clones and examined for RFLP. In 62 of them, polymorphism was detected with one or more enzymes; four RFLPs were VNTR systems. All polymorphic clones were assigned to one of 22 intervals obtained by mapping on a deletion panel of 15 somatic hybrid cell lines containing parts of chromosome 11; 11 clones were finely mapped by in situ hybridization. Although RFLP markers were scattered on the whole chromosome, they were found predominantly in the regions of R-banding. These DNA markers will contribute to fine mapping of genes causing inherited disorders and tumor-suppressor genes that reside on chromosome 11. Furthermore, as one-third of the cosmid clones revealed a band or bands in Chinese hamster DNA, indicating sequence conservation, this subset of clones may be useful for isolating biologically important genes on chromosome 11.